Abstract: In order to identify the peptides binding with different type of chicken MHC Ⅰ molecules, the constructed BF2*04sh-linker-β2m，BF2*09sh-linker-β2m，BF2*12sh-linker-β2m，BF2*05wj-linker-β2m proteins were mixed with seven designed and synthesized T cell epitopes (three derived from avian influenza virus, two derived from swine foot-and-mouth diseases virus and two derived from grass carp reovirus) at a respective mole ratio of 1∶10, incubated at 37℃ for 12 h. The mixtures were centrifuged in Amicon Ultra-15 protein Centrifugal Filter (3000) at 4 500 rpm for 45 min to remove unbound nonameric peptides. Then, the retained solution was mixed with 5 mL citrate-phosphate buffer, incubated at 37℃ for 5 min, then centrifuged as above to obtain the filtered solution.The peptides in acid elution solution were concentrated and desalted using Sep-Pak C₁₈ cartridges. The sample plate which obtained the lyophilized peptides dissolved in 10 μL of matrix solution was put into a 4700 Proteomic Analyzer, and the primary mass spectra (MS) and secondary spectra (MS/MS) were conducted.The results revealed that peptides KILTIYSTV and LLLAIVSLV which derived from avian influenza viruses (AIV) could bind with three different types of chicken MHC Ⅰ. The results confirmed that the binding ablity of different MHC Ⅰ with peptides are different, the T cell epitopes derived from different virus are MHC Ⅰ restricted. KILTIYSTV and LLLAIVSLV are avian influenza’s candidate T cell epitopes.